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The effect of the angiotensin II (AT 1A ) receptor stably transfected into human neuroblastoma SH-SY5Y cells on noradrenaline release and changes in intracellular calcium
A stable cell line expressing the angiotensin II (AII) receptor has been obtained by transfecting the human neuroblastoma SH-SY5Y with the plasmid pCEP4 containing the entire coding region of the rat angiotensin AII receptor AT 1A . Angiotensin II (AII; 1-100 nM) evokes the release of [ 3 H]noradrenaline ([ 3 H]NA) in this cell line. Pretreatment with 100 nM 12-O-tetradecanoylphorbol-13-acetate (TPA) enhances the AII-evoked release of [ 3 H]NA approximately two-fold. Removal of extracellular Ca 2+ ([Ca 2+ ] o ) decreases 100 nM AII-evoked release of [ 3 H]NA by over 50% both in the presence and absence of TPA. AII also increases intracellular Ca 2+ ([Ca 2+ ] i ) in this cell line which is consistent with the AT 1A receptor being coupled to phospholipase C. Pretreatment with 100 nM TPA for 8 min attenuated the effect of AII on [Ca 2+ ] i . The effects of AT 1A receptor stimulation are therefore regulated differently in this cell line by activation of protein kinase C (PKC). Thus a useful cell line has been obtained from the human neuroblastoma SH-SY5Y in which to study at the molecular level the mechanism(s) by which AII regulates NA release.